Glaucoma is a major cause of blindness worldwide. The number of cases where vision loss occurs is also increased by the rise in cases that go undiagnosed. The primary causes of this vision loss are the degeneration of the optic nerve and the reduction of retinal ganglion cells. Primary open-angle glaucoma (POAG) represents the most prevalent type of glaucoma, accounting for around two-thirds of all cases. Tears, aqueous humor (AH), and vitreous humor are three fluids in the eyes that contain extra cellular miRNAs. MicroRNAs (miRNAs) are naturally occurring non-coding RNAs and play vital roles in various physiological and pathological processes. The dysregulation of miRNAs in disease states can serve as a basis for biomarker identification. This study aimed to identify and re-evaluatekey miRNAs involved in POAG through a comprehensive analysis that included normalization, differential expression analysis, target gene identification, and Random Forest (RF) classification. Two public datasets were analyzed, which revealed both upregulated and downregulated miRNAs, and with subsequent target gene prediction, hsamiR-1298-5p was identified as a key miRNA with limited connectivity within the miRNet network. Enrichment analysis suggested potential processes involved in POAG, particularly the role of miR-9 in targeting VE-cadherin, leading to the reduction of β-catenin. Further exploration through Random Forest analysis identified important miRNAs, including hsamiR-1298, hsa-miR-574-5p, and hsa-miR-188-5p. These miRNAs have been studied for their involvement in the Wnt/β-caten in signaling pathway but not extensively in glaucoma, indicating a novel area for future research.